This Phase II SBIR project is focused at the development of a high-throughput screening method and instrument for drug discovery using miniaturized, cell-based assays. For developing new drugs against complex diseases such as diabetes, cancer and neurological and cardiovascular disorders, high throughput screening using cell-based assays will be essential. Miniaturization of cell-based assays for automation and reduced costs is required. GeneCard-HTS enables drug discovery using adherent cell-based assays. During the Phase I project, the feasibility of the GeneCard approach was demonstrated using apoptosis assays. A prototype instrument to automate GeneCard assay process was developed and assembled. During Phase II, the versatility and flexibility of the GeneCard platform will be investigated by performing other cell-based assays used in drug discovery. The proposed plan investigates the following three specific aims: ? 1. Build a high-throughput screening prototype system based on GeneCard technology (GeneCard-HTS). ? 2. Miniaturize multiple cell-based assays for GeneCard-HTS platform. We will miniaturize four commonly used drug discovery assays in the GeneCard-HTS platform, i) Apoptosis assays using the Mitosensor reagents, ii) cell-based kinase assays, iii) fluorescent cytotoxicity assays, and iv) gene expression profiling assays. We will measure the performance of these assays in GeneCard-HTS platform and compare with performance obtained at present. ? 3. Validate GeneCard-HTS by cell-based screening of compound libraries. In order to validate the Genecard-HTS system, we will perform screening of a 5,000 compound focused library using cell-based kinase and apoptosis assays. We will measure validation parameters such as system lifetime, operating temperature and humidity range, inter- and intra- assay variability, and day-to-day variability. ? ? Commercial applications of the proposed system include pharmaceutical and biotechnoiogy drug discovery. Further integration of the instrument based on design specifications prepared in Phase I will be performed. Screening of drug compound libraries to identify potential lead compounds will be performed. ? ?
